ヒト骨髄におけるNKおよびLAK前駆細胞の分化促進因子の研究

人骨髓NK、LAK祖细胞分化促进因子的研究

• 批准号: 62570534
• 负责人: 依田 安弘
• 金额: $ 0.96万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570534/
• 关键词: 骨髄; NK前駆細胞; NK細胞の分化; LAK細胞の分化; 分化促進因子

《目的》本研究ではインターロイキン2(1L2)依存性のヒト骨髄NK前駆細胞の分化促進因子を見出すことを目的とした. 《方法》健常人から骨髄を採取し, 比重遠心法により単核球を分離した. これらをナイロンウールカラムに通し粘着細胞を除去した後, 半赤血球ロゼット形成法, 単クローン抗体(T101, OKM1, OKT11, Leu111)+家克補体処理により成熟型TおよびNK細胞を除去した. 既報のごとくこれらの前処理細胞を組換え型1L2(γ1L2)添加イスコフDMEM培地に浮遊し培養した. この培養系に組換え型1L1(γ1L1)γインターフエロンα(γIFNα)rIFNγ, TNFをそれぞれ添加し, ^3Hチミジン摂取, NK活性および表面形質の発現におよぼす影響を観察した.《結果》(1)1L11〜100U/mlを培養系に添加すると培養7日目の^3Hチミジン摂取が1L1の濃度依存性に増加し100U/mlで最高となった. 第8日目のNK(K562)活性も1L1添加により増大した. 培養21日目にFACSIVにより表面形質を観察すると, 成熟型(NKH1^+CD16^+)NK細胞の比率は, 1L2単独群では5-10%であった. これに対し1L2+1L1添加群では成熟型が20-25%であり成熟型の増加を認めた. さらに1L1添加群では, CD2陽性率も増加傾向を示した. (2)上記の培養系にIFNαを添加すると^3Hチミジン摂取, NK活性発現ともに濃度依存性に抑制された. IFNα添加培養21日目では, NKH1^+細胞の比率は有意に低下したが, その中の成熟型の比率には変化なかった. γIFNα添加培養では, ^3H摂取, NK活性の発現に対する抑制効果はIFNαに比して少なく, NKH1^+細胞率にも著変なかった. (3)TNFの効果は, IFNαとほぼ同様であった.《結論》以上の結果から次の結論を得た. (1)1L1は, 骨髄NK前駆細胞の分化促進傾向を示したが, 全細胞の成熟を完成させるには他の要因が必要と考えられる. (2)IFNα, IFNγおよびTNFにはNK細胞の分化促進作用を認めない.

Objective: in this study, we found that there was no significant difference between the two groups in this study. Objective: in this study, we used the pro-cellular differentiation promoting factor (NK) of dependent bone marrow (NK) in this study. "methods" normal people do not have a bone sample, and a specific gravity method is used to separate the nuclear ball. After the adhesion cells were removed, hemierythrocytic antibodies (T101, OKM1, OKT11, Leu111) + carotene antibodies (T101, CD8, CD8, Leu111) + carotene antibodies (T101, TACs, NK cells) were removed. The cell tissue type 1L2 (γ 1L2) was added in the first half of the experiment. The DMEM floatation rate was increased. The system of 1L1 (γ 1L1) gamma IFN α (γ IFN α) rIFN γ, the addition of TNF, the extraction of ^ 3H, the surface morphology of active NK, and the effect of 100U/ml on the surface morphology. "results" (1) the system of 1L11~100U/ml was added for 7 days and the highest level of 1L1 dependence was obtained. On the 8th day, NK (K562) active 1L1 was added. On the 21st, the surface shape of FACSIV, the cell ratio of mature (NKH1 ^ + CD16 ^ +) NK, and the single group of 1L2 (5-10%). The mature type is 20-25%, the mature type is the same as the mature group, the 1L2+1L1 additive group is the mature group. The 1L1 adds a group of characters, and the CD2 sex rate increases to show you. (2) when IFN α was added to the culture system, the activity of NK was inhibited by concentration dependence. When IFN α is added on the 21st day, the ratio of NKH1 ^ + cells is intentionally low, while the ratio of mature cells in NKH1 α is low. When γ-IFN α was added to the culture medium, the activity of NK was inhibited. The activity of IFN α was significantly lower than that of NKH1. (3) the results of TNF were similar to those of IFN α. The results of the above results were similar to those of the previous results. (1) 1L1, the differentiation of precursor cells of bone NK promotes the development of cell differentiation, and the completion of whole cell maturation is due to the need for examination. (2) IFN α, IFN γ promoted the differentiation of TNF cells and NK cells.